co-ip - (Nov/02/2006 )

Hi everyone,

I have identified 6 proteins which interacts my gene of interest in the mating based yeast two hybrid sysytem. I would like to prove this interaction with CO-IP, di I need antibody for my gene of interest which I have used as my bait in the two hybrid system or do I need specific antibodies for the interactors or the preys which I have found, also is there any easier method to prove the two hybrid interaction ?...

---

You need an Ab to immunoprecipitate your protein of interest plus antibodies to detect the interactors. There are other methods to detect an interaction but the coIP is quite trivial.

---

in addition to dnafactory, ideally you need a second set of each Ab for Ib as it is not to recommend run Ib with the Ab´s for IP

---

i suggest that u fuse your bait protein or the protein of interest with a tag ie flag, myc, after that the ab could be bought from company.co-IP is such as:
the complex flag-A-B-myc can be pulled down by myc beads, so u can detect A with flag ab.

---